• Title/Summary/Keyword: Rac

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Actin-binding LIM protein 1 regulates receptor activator of NF-κB ligand-mediated osteoclast differentiation and motility

  • Jin, Su Hyun;Kim, Hyunsoo;Gu, Dong Ryun;Park, Keun Ha;Lee, Young Rae;Choi, Yongwon;Lee, Seoung Hoon
    • BMB Reports
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    • v.51 no.7
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    • pp.356-361
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    • 2018
  • Actin-binding LIM protein 1 (ABLIM1), a member of the LIM-domain protein family, mediates interactions between actin filaments and cytoplasmic targets. However, the role of ABLIM1 in osteoclast and bone metabolism has not been reported. In the present study, we investigated the role of ABLIM1 in the receptor activator of $NF-{\kappa}B$ ligand (RANKL)-mediated osteoclastogenesis. ABLIM1 expression was induced by RANKL treatment and knockdown of ABLIM1 by retrovirus infection containing Ablim1-specific short hairpin RNA (shAblim1) decreased mature osteoclast formation and bone resorption activity in a RANKL-dose dependent manner. Coincident with the downregulated expression of osteoclast differentiation marker genes, the expression levels of c-Fos and the nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), critical transcription factors of osteoclastogenesis, were also decreased in shAblim1-infected osteoclasts during RANKL-mediated osteoclast differentiation. In addition, the motility of preosteoclast was reduced by ABLIM1 knockdown via modulation of the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/Akt/Rac1 signaling pathway, suggesting another regulatory mechanism of ABLIM1 in osteoclast formation. These data demonstrated that ABLIM1 is a positive regulator of RANKL-mediated osteoclast formation via the modulation of the differentiation and PI3K/Akt/Rac1-dependent motility.

An Experimental Study to Determine the Mechanical Properties of Recycled Aggregate Separated from Demolished Concrete and Recycled Aggregate Concrete (폐 콘크리트에서 분리된 재생골재와 재생콘크리트의 공학적 특성규명을 위한 실험적 연구)

  • 전쌍순;이효민;황진연;진치섭;박현재
    • The Journal of Engineering Geology
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    • v.13 no.3
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    • pp.345-358
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    • 2003
  • Recently, the reuse of coarse aggregate derived from demolished concrete was introduced into practice with two environmental aspects: protection of natural sources of aggregate and recycling of construction waste. However, recycled aggregate has been used for the very limited application such as subbase material for pavement and constructional filling material because it was considered as low quality constructional materials. In the present study, in order to examine the possibility that recycled aggregate can be used for concrete mixing, we conducted various experimental tests to identify mineralogical, chemical and mechanical properties of recycled aggregate and to determine the workability and mechanical properties of recycled aggregate concrete (RAC). The cement paste and mortar contained in recycled aggregate significantly affect the basic mechanical properties of aggregate and the workability and mechanical properties of RAC. However, RCA mixed with the proper replacement ratio of recycled aggregate shows the comparable compressive strength and freeze and thaw resistance to those of normal concrete. Therefore, it is considered that recycled aggregate can be widely used for concrete if the cement paste and mortar can be efficiently removed from recycled aggregate and/or if the effective replacement ratios of recycled aggregate are applied for mixing concrete.

Reinforced fibrous recycled aggregate concrete element subjected to uniaxial tensile loading

  • Hameed, R.;Hasnain, K.;Riaz, M. Rizwan;Khan, Qasim S.;Siddiqi, Zahid A.
    • Advances in concrete construction
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    • v.9 no.2
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    • pp.195-205
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    • 2020
  • In this study, effect of recycled aggregates and polypropylene fibers on the response of conventionally reinforced concrete element subjected to tensile loading in terms of tension stiffening and strain development was experimentally investigated. For this purpose, concrete prisms of 100 × 100 mm cross section and 500 mm length having one central deformed steel re-bar were cast using fibrous and non-fibrous Recycled Aggregate Concrete (RAC) with varying percentages of recycled aggregates (0%, 25%, 50%, 75% and 100%) and tested under uniaxial tensile load. For all fibrous RAC mixes, polypropylene fibers were used at constant dosage of 3.15 kg/㎥. Effect of recycled aggregates and fibers on the compressive strength of concrete was also explored in this study. Through studying tensile load versus global axial deformation of composite and strain development in concrete and steel, it was found that replacement of natural aggregates with recycled aggregates in concrete negatively affected the cracking load, tension stiffening and strain development, and this negative effect was observed to be increased with increasing contents of recycled aggregates in concrete. The results of this study showed that it was possible to minimize the negative effect of recycled aggregates in concrete by the addition of polypropylene fibers. Reinforced concrete element constructed using concrete containing 50% recycled aggregates and polypropylene fibers exhibited cracking behavior, tension stiffening and strain development response almost similar to that of concrete element constructed using natural aggregate concrete without fiber.

Effects of Comonomer and Various Polymerization Conditions on Terpolymerization (삼원공중합에 있어서 공단량체 및 여러 가지 중합조건이 미치는 영향)

  • Kim, Jung Soo;Jeon, Dong-Gyu;Kim, Tae-Wan;Kim, Hyun Ki;Kim, Dong Hyun
    • Elastomers and Composites
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    • v.48 no.4
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    • pp.263-268
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    • 2013
  • In this study, we prepared poly(ethylene-ter-1-hexene-ter-divinylbenzene) using bridged rac-$Et[Ind]_2ZrCl_2$ metallocene catalysts. The effect of 1-hexene on the terpolymerization rate was evaluated. When cocatalyst/catalyst molar ratio was 3,000, catalytic activity indicated more than 8,000 which was very remarkable value. As polymerization time increased, the weight-average molecular weight of the terpolymer gradually increased to some degree. In case of a polymerization time of 50 minutes, the terpolymer became amorphous state. The molecular weight distribution and densities of the terpolymer were 110,000-200,000 and $0.85-0.89g/cm^3$, respectively. Thermal properties and structure of the terpolymer were also identified.

Cell Motility Is Decreased in Macrophages Activated by Cancer Cell-Conditioned Medium

  • Go, Ahreum;Ryu, Yun-Kyoung;Lee, Jae-Wook;Moon, Eun-Yi
    • Biomolecules & Therapeutics
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    • v.21 no.6
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    • pp.481-486
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    • 2013
  • Macrophages play a role in innate immune responses to various foreign antigens. Many products from primary tumors influence the activation and transmigration of macrophages. Here, we investigated a migration of macrophages stimulated with cancer cell culture-conditioned medium (CM). Macrophage activation by treatment with CM of B16F10 cells were judged by the increase in protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX2). The location where macrophages were at 4 h-incubation with control medium or CM was different from where they were at 5 h-incubation in culture dish. Percentage of superimposed macrophages at every 1 h interval was gradually increased by CM treatment as compared to control. Total coverage of migrated track expressed in coordinates was smaller and total distance of migration was shorter in CM-treated macrophages than that in control. Rac1 activity in CM-treated macrophages was also decreased as compared to that in control. When macrophages were treated with CM in the presence of dexamethasone (Dex), an increase in COX2 protein levels, and a decrease in Rac1 activity and total coverage of migration were reversed. In the meanwhile, biphasic changes were detected by Dex treatment in section distance of migration at each time interval, which was more decreased at early time and then increased at later time. Taken together, data demonstrate that macrophage motility could be reduced in accordance with activation in response to cancer cell products. It suggests that macrophage motility could be a novel marker to monitor cancer-associated inflammatory diseases and the efficacy of anti-inflammatory agents.

Copolymerization of Ethylene and Cycloolefin with Metallocene Catalyst: II. Effect of Cycloolefin (메탈로센 촉매를 이용한 에틸렌과 시클로올레핀의 공중합: II. 시클로올레핀의 영향)

  • 이동호;정희경;최이영;김현준;김우식
    • Polymer(Korea)
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    • v.24 no.6
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    • pp.751-756
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    • 2000
  • The copolymerization of ethylene (E) and cycloolefin (CO) was carried out with rac-Et(Ind)$_2$ZrC $l_2$ and MMAO cocatalyst system to examine the effect of CO structure on catalytic behaviors and properties of copolymer (COC). Various cycloolefins such as norbornene (N), 5-phenyl-2-norbornene (PN) and 5-vinyl-2-norbornene (VN) were used as comonomers. With increasing [CO]/[E] feed ratio, the catalytic activity decreased while the glass transition temperature of copolymer increased. With analysis of the structure of E/VN copolymer by FT-IR and $^{l3}$C-NMR, it was found that the cyclic C=C bond of VN comonomer is selectively polymerized and the vinyl C=C bond remains unreacted. The resulting vinyl C=C bond attached into copolymer provided the functionalization moiety using glycidyl methacrylate.e.

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Evaluation and cloning of a (R)-stereospecific esterase from Bacillus stearothermophilus JY144

  • Kim, Ji-Yeon;Kim, Yun-Jeong;Choe, Gi-Seop;Kim, Geun-Jung;Yu, Yeon-U
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.457-460
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    • 2002
  • In an effort to isolate novel strains expressing a thermostable esterase that hydrolyzed the rac-ketoprofen ethyl ester to ketoprofen in the stereospecific manner, we screened various soils and composts from broad ecological niches in which the activity was expected to be found. Three hundreds of microbial strains were tested to determine their ester-hydrolyzing activity by using an agar plate containing insoluble tributyrin as an indicative substrate, and then further screened by activity on the (R,S)-ketoprofen ethyl ester. Twenty-six strains were screened primarily at high growth and incubation temperature and further compared the ability to ethyl ester-hydrolyzing activity in terms of conversion yield and chiral specificity. Consequently, a strain JYl44 was isolated as a novel strain that produced a (R)-stereospecific esterase with high stability and systematically identified as a Bacillus stearothermophilus JY144. The enzyme indeed stables at a broad range of temperature, upto 65 $^{\circ}C$, and pH ranging from 6.0 to 10.0. The optimal temperature and pH for enzymatic conversion were 50 $^{\circ}C$ and 9.0, respectively. Based on the observations that resulted a poor cell growth, and enzyme expression in wild type strain, we further attempted the gene cloning into a general host Escherichia coli and determined its primary structure, concomitantly resulting a high level expression of the enzyme. The cloned gene had an open reading frame (250 amino acids) with a calculated molecular mass of 27.4 kDa, and its primary structure showed a relative high homology (45-52 %) to the esterases from Streptomyces and Bacillus strains. The recombinant whole cell enzyme could efficiently convert the rac-ketoprofen ethyl ester to (R)-ketoprofen, with optical purity of 99 % and yield of 49 %.

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A Study on Single Cell Polarized Signals Using Polydimethylsiloxane-based Micropatterned Channel System (폴리디메틸실록산 기반 마이크로패턴 채널 시스템을 이용한 단일 세포의 극성 신호에 관한 연구)

  • Suh, Jung-Soo;Lee, Chanbin;Pan, Yijia;Wang, Yingxiao;Jung, Youngmi;Kim, Tae-Jin
    • Korean Chemical Engineering Research
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    • v.58 no.1
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    • pp.122-126
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    • 2020
  • In this study, we produced the micropatterned channel system using polydimethylsiloxane (PDMS) and micromolding in capillaries (MIMIC) technology and evaluated cellular polarity signals through high-resolved imaging at the single-cell level. In cells treated with platelet-derived growth factor (PDGF), three types of key signals in cell migration; phosphoinositide 3-kinase (PI3 K), Rac, and Actin, were strongly activated in the front area compared to the rear region, whereas myosin light chain (MLC) showed no notable activity in the front and rear areas. Our results will, therefore, provide important information and methodology for studying the correlation between cell polarity signals and cell migration under the newly defined microenvironment.

All-trans Retinoic Acid-Associated Low Molecular Weight Water-Soluble Chitosan N anoparticles Based on Ion Complex

  • Kim Dong-Gon;Choi Changyong;Jeong Young-Il;Jang Mi-Kyeong;Nah Jae-Woon;Kang Seong-Koo;Bang Moon-Soo
    • Macromolecular Research
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    • v.14 no.1
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    • pp.66-72
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    • 2006
  • The purpose of this study is to develop novel nanoparticles based on polyion complex formation between low molecular weight water-soluble chitosan (LMWSC) and all-trans retinoic acid (atRA). LMWSC nanoparticles encapsulating atRA based on polyion complex were prepared by mixing of atRA into LMWSC aqueous solution using ultrasonication. In FTIR spectra, the carbonyl group of atRA at 1690 $cm^{-1}$ disappeared or decreased when ion complexes were formed between LMWSC and atRA. In ${1}^H$ NMR spectra, specific peaks of atRA disappeared when atRA-encapsulated LMWSC (RAC) nanoparticles were reconstituted into $D_{2}O$ while specific peaks both of atRA and LMWSC appeared in $D_{2}O$/DMSO (1/3, v/v) mixture. XRD patterns also showed that the crystal peaks of atRA were disappeared by encapsulation into LMWSC nanoparticles. LMWSC nanoparticles encapsulating atRA have spherical shapes with particle size below 200 nm. The mechanism of encapsulation of atRA into LMWSC nanoparticles was thought to be an ion complex formation between LMWSC and atRA. LMWSC nanoparticles showed high atRA loading efficiency over 90$\%$ (w/w). AtRA was continuously released from nanoparticles over 10 days. In in vitro cell cytotoxicity test, free atRA showed higher cytotoxic effect against CT 26 colon carcinoma cell line on 1 day. However, RAC nanoparticles showed similar cytotoxicity against CT 26 cells on 2 day. These results suggest the potential for the introduction of LMWSC nanoparticles into various biomedical fields such as drug delivery.

Entamoeba histolytica Induces Cell Death of HT29 Colonic Epithelial Cells via NOX1-Derived ROS

  • Kim, Kyeong Ah;Kim, Ju Young;Lee, Young Ah;Min, Arim;Bahk, Young Yil;Shin, Myeong Heon
    • Parasites, Hosts and Diseases
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    • v.51 no.1
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    • pp.61-68
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    • 2013
  • Entamoeba histolytica, which causes amoebic colitis and occasionally liver abscess in humans, is able to induce host cell death. However, signaling mechanisms of colon cell death induced by E. histolytica are not fully elucidated. In this study, we investigated the signaling role of NOX in cell death of HT29 colonic epithelial cells induced by E. histolytica. Incubation of HT29 cells with amoebic trophozoites resulted in DNA fragmentation that is a hallmark of apoptotic cell death. In addition, E. histolytica generate intracellular reactive oxygen species (ROS) in a contact-dependent manner. Inhibition of intracellular ROS level with treatment with DPI, an inhibitor of NADPH oxidases (NOXs), decreased Entamoebainduced ROS generation and cell death in HT29 cells. However, pan-caspase inhibitor did not affect E. histolytica-induced HT29 cell death. In HT29 cells, catalytic subunit NOX1 and regulatory subunit Rac1 for NOX1 activation were highly expressed. We next investigated whether NADPH oxidase 1 (NOX1)-derived ROS is closely associated with HT29 cell death induced by E. histolytica. Suppression of Rac1 by siRNA significantly inhibited Entamoeba-induced cell death. Moreover, knockdown of NOX1 by siRNA, effectively inhibited E. histolytica-triggered DNA fragmentation in HT29 cells. These results suggest that NOX1-derived ROS is required for apoptotic cell death in HT29 colon epithelial cells induced by E. histolytica.